Experiment Study Of Effect Of Tongfengning On Cyclooxygenase And Cytokines In Gouty Arthritis Synovial Cells

Objective To further investigate the inflammation mechanism in gouty arthritis and efficacy of Tongfengning (TFN) for its anti-inflammation and pain-relief by observing cyclooxygenase (COX) isoforms, mterleukin-1 (IL-1) and tumor necrosis factor (TNF ) in monosodium urate (MSU) crystals stimulated synovial cells in vivo and vitro.Methods (1)Rats were randomly divided into four groups: Control, Model, Chinese Herb (TFN) and Medicine (Colchicine) Group. The former two groups were treated with distilled water and the latter two with TFN or colchicine solution respectively. After 5 days, the acute gout models were established by injecting MSU solution into rat ankles except Control Group injected saline instead. Biopsies were taken from ankle synovium in Control, TFN and Colchicine Group after 6h and Model Group in different time (after 1h, 3h, 6h, 12h, 24h, 48h individual) . The expressions of COX-1/COX-2 in rat synovium were detected with immunohistochemical method of SP. (2)The rat cultured synovial cells in vitro were mixed with the different concentration of MSU solution and incubated for 24h. COX-1/COX-2 production, protein and mRNA of IL-1 and TNF were quantitated with ELISA and RT-PCR. Similar method of detection was administered in the rat cultured synovial cells co-incubated with 0.16mg/ml MSU solution and the different diluted serum containing TFN.Results (3)The expression of COX-l/COX-2 in synovial cells peaked in 3-6h in Model Group after MSU injected (P<0.01), especially COX-2. In othertime periods, the expression of COX-2 , not COX-1, also increased significantly. TFN and colchicine could downregulate the expression of COX-l/COX-2 in gouty arthritis synovial cells with no significant difference between them. (2) Cultured synovial cells induced by MSU crystals showed high level COX-l/COX-2 production (P<0.01), especially for COX-2. IL-1 , TNF mRNA and protein production increased correspondingly and have a positive correlation with the concentration of MSU crystals. COX-l/COX-2 production could be inhibited by the serum containing TFN (P<0.01), especially for COX-2. IL- 1, TNF mKNA and protein also demonstrated a significant decrease which correlated with the diluted rate of serum containing TFN in negative way.Conclusions Synovial cells in acute gout showed a significant increase in the expression of COX-l/COX-2 with a high level in transcript and translation of IL-1 , TNF accordingly. TFN might have a anti-inflammation and pain-relief effect on gouty arthritis synovial cells through inhibiting productions of COX-l/COX-2 and proinfiarnmation cytokines such as IL-lp, TNFa.